抗人载脂蛋白B单克隆抗体杂交瘤细胞株的建立及应用

PREPARATION AND APPLICATION OF MOUSE HYDRIDOMA CELL LINE WITH SECRETING MONOCLONAL ANTIBODY OF HUMAN APOLIPOPROTEINB

用人血清分离纯化的载脂蛋白B(Apo B)作为抗原免疫BaLE/C小鼠,取脾脏制成脾细胞悬液与SP2/O小鼠骨髓瘤细胞融合,融合率为19.3％。融合后检测抗体,经第二次亚克隆培养,抗体阳性率已达100％。杂交瘤细胞上清液效价为1:256,腹水效价为8×10^(-6),经聚丙烯酰胺凝胶电泳鉴定,此纯化的单克隆抗体显示单一区带。经免疫电泳和免疫扩散法证实此免疫球蛋白属IgG2b。与ApoA_Ⅰ、A_Ⅱ、Cs、E及人血清白蛋白均无交叉反应。比较用单克隆抗体法与浊度法测定血浆ApoB水平的结果时,发现它们之间呈显著相关性(r=0.85)。

A purified apolipoprotein B as an antigen was injected into female Balb/c mice subcutaneously. All hydridoma were prepared by fusing spleen cells of immunized mice treated as above with a non-secreting myeloma cells line SP2-O. The positive fusing rate was 19.3%. After secondary incubation of subcloning, The antibody positive reaction reached 100%. The vallence of supernatant of hydridoma cells and ascites were 1:256 and 8x 10^(-6), respectivety. The immunoglobin class of each antibody was determined and confirmed by immunoelectrophoresis and immunodifusion techniques as IgG2b subclass. No immunocross reaction with ApoAⅠ, ApoAⅡ, ApoCs and albumin had been detected. The purified monoclonal antibody showed a single band on polyacrymide gel electrophoresis. When compared the results obtained from two methods i.e. monoclonal antibody technique and nephelometry in determining the plasma ApoB level, a significante correlation (r=0.85) had been observed.